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Polyphenols, like flavonoids, have been investigated when present in intact plants or in extracts as methane mitigating dietary supplements in ruminants. The aim of the present study was to examine pure compounds in a short-term in vitro experiment using the Hohenheim Gas Test method. We focused on the group of the flavonoids and tested which of them had the potential to mitigate methane without negatively affecting ruminal fermentation. Eight flavonoids were tested: epicatechin, luteolin-7-glucoside, quercetin, and isoquercetin in Experiment 1; catechin, gallocatechin, epigallocatechin, and epigallocatechin gallate in Experiment 2. Tannic acid, no flavonoid but a phenolic acid with known methane mitigating properties, served as positive control, and the unsupplemented basal diet as negative control. In both experiments, each of these compounds (including tannic acid) was tested at dosages of 0.5, 5.0, and 50.0 mg/g basal diet dry matter (DM) in four runs each. Gallocatechin, tannic acid, and epigallocatechin gallate (50 mg/g DM) lowered fermentation gas formation and in vitro organic matter digestibility relative to the negative control (Experiment 2). Apart from tannic acid, epicatechin, quercetin, isoquercetin, and luteolin-7-glucoside (5 and 50 mg/g DM) reduced the amount of CH4 produced in relation to total gas produced (Experiment 1). The incubation fluid ammonia concentration was decreased with luteolin-7-glucoside and tannic acid (50 mg/g DM). From the flavonoids tested especially luteolin-7-glucoside seems to have a similar potential as tannic acid to mitigate methane and ammonia formation during ruminal fermentation in vitro, both favourable in environmental respect. These results need to be confirmed in live animals.

In vitro bioactivity of various pure flavonoids in ruminal fermentation, with special reference to methane formation